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Measurement of type I interferon production

Rosalind E Seeds1, Joanna L Miller

  • 1Sir William Dunn School of Pathology, University of Oxford, United Kingdom.

Current Protocols in Immunology
|March 15, 2011
PubMed

Insights

This study presents multiple methods for measuring murine interferon (IFN)α/β, including flow cytometry, ELISA, and bioassays. These complementary techniques enable comprehensive assessment of IFN production and antiviral activity in mouse cells.

Area of Science:

  • Immunology
  • Virology
  • Cell Biology

Background:

  • Interferons (IFNs) are crucial cytokines in the innate immune response against viral infections.
  • Accurate measurement of IFN production is essential for understanding immune responses and developing antiviral therapies.
  • Murine models are frequently used to study IFN function and antiviral immunity.

Purpose of the Study:

  • To describe and compare various methods for quantifying murine interferon (IFN)α/β.
  • To provide protocols for intracellular cytokine staining, ELISA, and biological assays for IFN measurement.
  • To enable researchers to select the most appropriate assay for their specific needs in studying murine IFN production.

Main Methods:

  • Intracellular staining for IFNα/β followed by flow cytometry (FACS assay).
  • Enzyme-linked immunosorbent assay (ELISA) for quantifying secreted IFNα.
  • Biological assay to measure the functional antiviral activity of IFN.

Main Results:

  • The FACS assay allows for the measurement of IFNα/β production within defined murine cell populations.
  • ELISA quantifies the amount of secreted IFNα.
  • The biological assay is the most sensitive method, measuring functional antiviral activity.

Conclusions:

  • A combination of flow cytometry, ELISA, and biological assays provides complementary approaches for comprehensive IFN assessment in murine systems.
  • These methods facilitate the detailed characterization of IFNα/β production and function.
  • The choice of assay depends on whether intracellular or secreted IFN, or functional antiviral activity, needs to be measured.

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